Part:BBa_K4099008

pIB165

Usage and Biology

The plasmid pIB165 is a E. coli-Streptococci shuttle plasmid for gene expression in streptococci with P23 promoter.

Experimental approach

We extracted the pIB165 plasmid from E. coli for later electrotransformation. Its sequence map is shown in Figure 1.

Figure 1. DNA profile of the plasmid pIB165..

Proof of function

After we electrotransformed the plasmid pIB165 as the foreign DNA to test the transformation efficiency of our modified L. casei, it took several days to culture and finally saw the comparison results. As showing above, we can see that the modified L. casei (KO) has higher transformation efficiency with remarkably more colonies than the wild (Wild).

Transformation test result

Figure 2. Comparison between the wild L. casei (left) and modified L. casei (right, LSEI-2094 knocked out) in transformation..

Graph 1. Comparison between the wild L. casei and modified L. casei in transformation

Figure 3. Histogram comparison between wild strain and KO strain..

In addition, we measured OD600 of these strain groups which were pre-spreaded plates with different volumes of bacteria solutions so as to quantify the transformation results as showing above (Fig. 3). In conclusion, it indicates that our modified L. casei has much higher efficiency of the foreign plasmid transformation than the wild and the modified L. casei has great potential to be used as the recombinant carrier in various areas.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 3991
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 4066
  Illegal XhoI site found at 4072
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]